PROJECT SUMMARY/ABSTRACT Chronic inflammation, defined by a persistent elevation of local and systemic pro-inflammatory factors, is a hallmark of biological aging. We and others have shown that chronic levels of circulating downstream markers of systemic inflammation are associated with age-related disease states, particularly cardiovascular disease, as well as longevity. However, accumulating evidence suggests that upstream mediators of inflammation are more likely to play a causal role in disease pathogenesis and, in turn, serve as effective therapeutic targets. Upstream initiation of inflammation in humans is governed primarily by small molecule effectors of arachidonic acid metabolism, termed eicosanoids. These bioactive lipid effectors of both pro- and anti-inflammatory activity include thromboxanes, prostaglandins, lipoxins, and leukotrienes. To date, the interaction between eicosanoid pathways and age-related disease phenotypes remain poorly understood, thus limiting our ability to harness their therapeutic potential. This proposal aims to provide a more detailed understanding of how upstream eicosanoid pathways can be variably active, imbalanced, and perturbed in relation to an individual?s propensity for developing or escaping age-related disease. Emerging data suggest that select eicosanoids and genetic polymorphisms in enzymes contributing to their biosynthesis are associated particularly with cardiovascular risk as well as longevity, but comprehensive studies in large cohorts are lacking. Advanced methods using mass spectrometry now allow for the rapid and accurate quantification of >150 upstream eicosanoid mediators representing the multiple enzymatic origins. We will use these methods to comprehensively assay distinct pro- and anti-inflammatory eicosanoids and examine their relation to measures of healthy cardiovascular aging and longevity outcomes in a large cohort of women, enriched with individuals representing the spectrum of risk factor burden as well as a range of achieved lifespans. The goal of this study is to advance our understanding of how upstream inflammatory pathways are related to healthy aging and longevity in women, who continue to outlive men and yet carry a greater burden of chronic disease in later life than men for reasons that remain unclear. Thus, our specific aims are (1) to assess whether circulating eicosanoid mediators of systemic inflammation are associated with longevity in women; and, (2) to evaluate the association between distinct eicosanoid mediators of systemic inflammation and morbidity profiles in women. Our systematic approach to comprehensively investigating the components of upstream inflammatory activity in a large, well-phenotyped cohort promises to yield important insights into the determinants of healthy aging and longevity. Importantly, given its focus on upstream inflammatory activity, this work will pave the way for follow-up studies investigating the efficacy of anti-inflammatory therapies (both existing and novel agents) for modulating variation in distinct eicosanoids as well as outcomes.